[This message has been edited by joee (edited November 30, 1999).]

I have had some success using H2O2 in the sterilized water that the mycelium is blended with to make mycelium water. I do this when I make up fresh "shroom water" to clone a fruitbody. I pretty much follow 9'ers tek for this, except I add 2ml 3%H2O2 per 100ml H2O after the water has cooled and just before the chunk of mycelium is added and blended. I add the mycelium (usually a skinned piece of a stem) and give it 2, four second bursts of the blender to shred and distribute the mycelium.

This "peroxidated" shroom water will foam up a little showing that the peroxide is immediately broken down. My reasons for doing this because:
1. It helps make sure that contaminates aren't introduced with the chunk of mycelium tissue, but are hopefully killed immediately during blending. I rarely see any contamination at all using this, even though I start with a non-sterile chunk of stem.

2. It should help prevent the germination of any mushroom spores that might be lodged in the chunk of tissue. When I do this I am generally trying to generate clones of a specific mushroom. I don't want spores that might germinate and create a multi-strain culture to get carried along. Stems usually have visible spores on them. I don't know if this idea is entirely successful.

I inoculate with the shroom water within 20 minutes after it is made up, I don't think it would store for long as the mycelium is extra stressed by the peroxide. I use standard PF type syringe inoculations of sterilized jars. I usually use a "massive" 10cc per 1/2 pint which really gets the jars going fast. Massive inoculation with cloned mycelium water usually cuts in 1/2 the time to grow out jars compared to spore inoculation.

I have noticed at the concentration above that the mycelium is definately stressed a little. It is important to blend it into no smaller pieces than absolutely necessary, i.e. what will barely fit up the inoculation needle without clogging too badly.

Different species seem to have different abilities to withstand this peroxide treatment. Cubensis isn't stressed much at all. Ps. cyanescens and azurescens both do well. Shiitake and oysters both do well. Ps. tampanensis and Portabellas do ok, but are a little more delicate. Panaeolus cyanescens and Ps. semilanceata are both more delicate and can be killed unless you reduce the concentration.

I don't know if the above answered your question or not. I don't use non-sterile bulk substrates so I don't know if they would work with peroxide and mycelium water inoculation or not. I suspect you would have problems though because of the longer exposure and greater quantity of peroxide the mycelium must endure while it is in a delicate state. That's just a guess though.

Joee, I think you got it backwards. Its not the mycelium that can't stand up to the h2o2, its the peroxide. The mycelium in mycelium water quickly breaks down the h2o2. It can be used, as the doc pointed out, but it can't be used in substitute for sterile teks which is what peroxide man is aiming for.

Using mycelium water on peroxidated substrates has a similar effect. Mycelium is spread out throughout the substrate, which is what is good about using it. This however, breaks down all the peroxide in the substrate. In the manual, for example, Wayne suggests using agar wedges to innoc peroxide subs, and not shaking untill there is a good bit of growth. By doing this, the peroxide that isn't near the point of innoculation stays active until shaking. After shaking, colonization continues so rapidly that the peroxide isn't needed anymore.

I think that's what you were looking for. The Dr is right. You can use peroxide in both places with myc water, but it's help isn't as pronounced as with other peroxide methods, such as agar.

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-From a registered Mad Scientist

"From a certain point of view"
-Jedi Master Obiwan Kenobi (also a Mad Scientist tm)

[This message has been edited by mycofile (edited November 30, 1999).]

Great Tek, how do you blend the piece of schroom, In the blender (if how do sterize it?), in a jar (how you blend it in there)

sounds quite easy, and I wouldn`t need agar anymore ( to seperate a single strain)

thanx

hate to post dump questions, but sorry i did it....

[This message has been edited by Prellgott (edited November 30, 1999).]

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It?s easy to do without silver and gold
or even pleasures of love
but its hard to do without mushrooms.

Marcus Valerius Martialis
Roman poet

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-From a registered Mad Scientist

"From a certain point of view"
-Jedi Master Obiwan Kenobi (also a Mad Scientist tm)